Inhibition of EMT in colorectal cancer by rosemarinic acid explored based on the Wnt/β-catenin pathway
Objective To investigate the role and action mechanism of rosemarinic acid(RA)on the malignant behavior of colorectal cancer(CRC)SW480 cells.Methods We collected tumor tissues from patients with primary untreated CRC who underwent surgery in People's Hospital of Rugao from January 2019 to December 2020,as well as colon tissues from normal individuals.The expression of β-catenin was detected by immunohistochemistry and Western blotting to analyze its relationship with the clinical stage,prognosis,and immune infiltration.CCK-8 was used to detect the effects of 10,15 and 20 μmol/L of RA on the proliferation of SW480 cells;flow cytometry detected apoptosis;Transwell and scratch assay detected changes in the invasion and migration ability of the cells,respectively.Western blotting detected the expressions of proteins related to apoptosis,epithelial-mesenchymal transition(EMT),and Wnt/β-catenin pathway.A mouse model of CRC transplant tumor was established,and the effect of RA on the growth of transplant tumor was investigated by gavage administration.The levels of inflammatory factors were detected by ELISA.Results The expression of β-catenin was low in normal colon tissues and significantly higher in CRC tissues.In the CRC patients with high expression of β-catenin,the number of patients with tumors larger than 5 cm was significantly higher than that in the low expression group,whereas the overall survival of the patients was significantly smaller than that in the low expression group(P＜0.05).Cell proliferation was significantly inhibited and decreased in a concentration-dependent manner in the RA group at all concentrations(P＜0.05).In animal experiments,the expression of apoptotic proteins Bax and Bad was significantly increased and the expression of anti-apoptotic protein Bcl-2 was significantly decreased in the RA-treated group compared with the control group(P＜0.05).It was also found that the invasive and migratory ability of SW480 cells was significantly inhibited(P＜0.05),the expression of E-cadherin protein was significantly increased,the expression of Snail,N-cadherin and Vimentin was significantly reduced(P＜0.05),the expression of Axin and GSK-3β was increased,and the expression of β-catenin was reduced(P＜0.05).Conclusion RA may inhibit the biological activity of SW480 by interfering with Wnt/β-catenin pathway-mediated EMT.
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